Following incubation with the reagents cells must be washed to reduce the possibility of background fluorescence from unspecific extracellular binding of ethidium homodimer or hydrolysis in aqueous solution of AM calcein.

In brief, patients with HER2 gene amplification by FISH and low HER2 total protein expression or homodimer levels as measured by the VeraTag assay [31] were found to respond poorly to trastuzumab-containing therapy, with Cox proportional hazards multivariate regression identifying HER2-HER2 dimer status as an independent predictor of time to progression (HR=0.

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